CellSorts CS-0026 Mouse CD3+ T Cell Sorting Kit — column-free negative selection, 90-99% purity
CS-0026

Mouse CD3+ T Cell Sorting Kit

Column-Free Negative Mouse

The Cell Sorts CS-0026 Mouse CD3+ T Cell Sorting Kit uses column-free negative selection to isolate untouched CD3+ T cells from mouse splenocytes and lymph node suspensions with 90–99% purity in 18–20 minutes. A biotin-conjugated antibody cocktail labels non-target cells (B cells, NK cells, monocytes, granulocytes), and streptavidin nanobeads bind them for magnetic removal — leaving untouched CD3+ T cells in their native state.

Compared to density gradient centrifugation (40–60% purity) or FACS (expensive, low throughput), this kit delivers 90–99% CD3+ purity with >95% recovery and viability. The streamlined column-free protocol requires no columns or centrifugation. Full preservation of TCR and CD3 signaling — ideal for CAR-T development, T cell activation, cytokine profiling, and tumor immunology research.

Sorting Purity
Up to 99%
Sorting Time
18–20 min

Product Features of the Mouse CD3+ T Cell Sorting Kit

The Mouse CD3+ T Cell Sorting Kit delivers industry-leading performance for rapid CD3+ T Cell isolation from PBMCs via column-free negative selection.

Column-free separation — No magnetic column needed; streamlined workflow isolates untouched CD3+ T cells directly from mouse splenocytes and lymph node cell suspensions by simple pour-off collection
Zero contact with target cells — Antibodies and streptavidin nanobeads bind only non-CD3+ cells; CD3+ T cells remain completely untouched in their native, unactivated state
Purity 90-99% — High-efficiency negative selection delivers ultra-pure CD3+ T cells populations, validated by flow cytometry
18-20 min fast protocol — Rapid column-free isolation from sample to purified CD3+ T cells, saving valuable research time
No bead residue — Isolated CD3+ T cells are completely bead-free and ready for immediate downstream applications including flow cytometry, cell culture, functional assays, CRISPR gene editing, scRNA-seq, TCR repertoire sequencing
Easy handling — Minimal steps, user-friendly procedure suitable for any lab; no expensive FACS equipment or specialized training required

The Mouse CD3+ T Cell Sorting Kit uses column-free negative selection to isolate untouched CD3+ T Cells from human PBMCs. To achieve these results, follow this simple 4-step protocol that takes just 18–20 min from start to finish. No centrifugation and no lengthy incubations. Furthermore, this streamlined protocol ensures consistent results across experiments.

How the Mouse CD3+ T Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

1
Step 1 of CD3+ T cells sorting: Sample Preparation - Prepare single-cell suspension from mouse tissue for CS-0026 Mouse CD3+ T Cell Sorting Kit

Step 1: Sample Preparation

Prepare single-cell suspension from mouse splenocytes and lymph node cell suspensions. Pass through 70 μm strainer, ACK lysis if needed. Count cells, resuspend at 1×10⁸ cells/mL in separation buffer (2–8°C).

2
Step 2 of CD3+ T cells sorting: Antibody Cocktail Labeling - Biotin-conjugated antibodies target non-CD3+ cells for depletion with CS-0026

Step 2: Antibody Cocktail Labeling

Add 10 μL Mouse CD3+ T Cell Isolation Cocktail per 1×10⁷ cells. Mix, incubate 4°C 5–10 min. Labels non-CD3+ cells (B cells, NK cells, monocytes, granulocytes, erythrocytes); CD3+ T cells untouched.

3
Step 3 of CD3+ T cells sorting: Magnetic Nanobead Binding - Streptavidin nanobeads bind labeled non-target cells for magnetic removal with CS-0026

Step 3: Magnetic Nanobead Binding

Vortex Streptavidin Nanobeads, add 10 μL per 1×10⁷ cells. Mix, incubate 4°C 5–10 min. Nanobeads bind labeled non-target cells.

4
Step 4 of CD3+ T cells sorting: Column-Free Separation - Purified CD3+ T cells collected by magnetic separator pour-off for CS-0026

Step 4: Column-Free Separation & Collection

Dilute to 2.5 mL, place on magnetic separator 3–5 min. Pour off supernatant containing purified CD3+ T cells — ready for T cell activation, cytokine profiling, CAR-T development, adoptive cell therapy, tumor immunology.

Applications for the Mouse CD3+ T Cell Sorting Kit

This Mouse CD3+ T Cell Sorting Kit supports a wide range of downstream applications for CD3+ T Cell isolation and column-free negative selection workflows from mouse splenocytes or lymph node cells.

Immunology Research — T cell activation, proliferation, and cytokine production studies in mouse models Autoimmune Disease Studies — EAE, colitis, and other mouse autoimmune model research Infectious Disease Research — Virus-specific T cell response analysis in mouse infection models Cancer Immunology — Tumor-infiltrating lymphocyte (TIL) studies and tumor microenvironment research Flow Cytometry — High-purity CD3 T cell populations ready for staining and analysis Cell Culture — Unactivated, bead-free T cells ideal for in vitro expansion and manipulation Drug Discovery — T cell-targeted drug screening in syngeneic and transgenic mouse models Adoptive Transfer — Highly viable, naive-state T cells for mouse adoptive cell transfer experiments

In addition, the Mouse CD3+ T Cell Sorting Kit is compatible with a wide range of downstream applications, making it a versatile tool for your immunology research workflow.

Kit Components

Each Mouse CD3+ T Cell Sorting Kit contains everything needed for high-purity column-free negative selection from human PBMCs.

  • Mouse CD3 T Cell Isolation Cocktail
  • Streptavidin Nanobeads
  • User manual

Storage & Shelf Life

Storage
Store at 2-8°C protected from light
Shelf Life
12 months

Mouse CD3+ T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0026 Mouse CD3+ T Cell Sorting Kit

Click each question to expand the answer

The CS-0026 Mouse CD3+ T Cell Sorting Kit from Cell Sorts uses column-free negative selection. Non-target cells (B cells, NK cells, monocytes, granulocytes, erythrocytes) are labeled with a biotin-conjugated antibody cocktail and removed by magnetic separation, while untouched CD3+ T cells remain in the supernatant. This ensures no beads or antibodies ever contact the target cells, preserving their native functional state.
Purity of isolated CD3+ T cells typically reaches 90-99%, as validated by flow cytometry using CD3-APC (Clone 17A2), CD4-FITC, CD8a-PerCP-Cy5.5 staining. The optimized antibody cocktail efficiently depletes all non-target cell populations while maximizing recovery of viable, functional CD3+ T cells from mouse splenocytes and lymph node cell suspensions.
The CS-0026 kit is optimized for mouse splenocytes and lymph node cell suspensions. Prepare a single-cell suspension by gentle mechanical disruption through a 70 μm cell strainer. Perform red blood cell lysis if needed (e.g., ACK lysis buffer). The kit is compatible with C57BL/6, BALB/c and other common mouse strains used in immunology research.
Step 1: Prepare a single-cell suspension from mouse splenocytes and lymph node cell suspensions.
Step 2: Count cells and resuspend at recommended density.
Step 3: Incubate with Mouse CD3+ T Cell Isolation Cocktail at 4°C for 5–10 min.
Step 4: Add Streptavidin Nanobeads and incubate at 4°C for 5–10 min.
Step 5: Place on magnetic separator for 3–5 min.
Step 6: Pour off purified CD3+ T cells into collection tube.
Total time: 18-20 min.
Isolated CD3+ T cells are immediately ready for: T cell activation, cytokine profiling, CAR-T development, adoptive cell therapy, tumor immunology, flow cytometry analysis, cell culture and expansion, CRISPR gene editing, single-cell RNA sequencing (scRNA-seq), and adoptive cell therapy studies. Zero bead or antibody interference means no additional washing steps are required.
Antibodies and streptavidin nanobeads bind exclusively to non-target cells—they never touch CD3+ T cells. This untouched approach ensures full preservation of surface receptors, native functional states, and physiological relevance for downstream functional assays. Cell Sorts column-free technology eliminates column-induced mechanical stress, further enhancing cell viability.
The antibody cocktail targets CD19, CD11b, CD49b, Ly-6G, Ter-119 on non-target cells—effectively labeling B cells, NK cells, monocytes, granulocytes, erythrocytes for depletion while leaving CD3+ T cells completely untouched. This comprehensive marker panel ensures high purity and recovery of the target cell population.
We recommend flow cytometry using CD3-APC (Clone 17A2), CD4-FITC, CD8a-PerCP-Cy5.5 antibodies. Typical results show 90-99% target cells in the collected fraction. Viability can be assessed using Trypan blue exclusion or 7-AAD staining. For best results, gate on lymphocytes by forward and side scatter, then analyze marker expression within the gated population.
Absolutely. Since isolated CD3+ T cells are completely bead-free and antibody-free, they are immediately compatible with single-cell RNA sequencing (scRNA-seq) and other single-cell technologies without additional washing steps. Cell Sorts protocols are validated for 10x Genomics and other major scRNA-seq platforms.
The CS-0026 uses column-free negative selection (18-20 min), ideal for speed and convenience with smaller samples. Column-based kits (25–35 min) offer higher consistency for larger sample sizes. Both yield untouched, bead-free CD3+ T cells with 90-99% purity. Choose CS-0026 for rapid processing; choose column-based for larger-scale isolation.

Related topics covered:

column-free negative selection CD3+ T Cell isolation CD3+ T Cell purity verification CD3+ T Cell sorting protocol column-free isolation single-cell analysis (scRNA-seq) CD3+ T Cell isolation markers PBMC CD3+ T Cell sorting

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